This application claims priority from Japanese Application No. 2000-147116, filed May 18, 2000, and Japanese Application No. 2001-99508, filed Mar. 30, 2001.
The present invention relates to human antibodies which bind to AILIM (activation inducible lymphocyte immunomodulatory molecule, also referred to as ICOS (inducible co-stimulator); human monoclonal antibodies which bind to AILIM or a portion thereof; DNA encoding said human monoclonal antibody or a portion thereof, or a portion of said DNA; cells (including genetic recombinant cells) producing said human monoclonal antibody or a portion thereof; human monoclonal antibody or a portion thereof produced by said genetic recombinant cells; pharmaceutical composition comprising said human monoclonal antibody or a portion thereof; pharmaceutical composition comprising antibody to AILIM for treating disorders related to the delayed allergy; method for identifying, quantitating or assaying substances that bind to AILIM or AILIM ligand; and kit used for said method.
A living body of mammals has immune response systems that excludes pathogenic microorganisms (viruses, bacteria, parasites, etc.) or foreign bodies (both are called xe2x80x9cantigenxe2x80x9d in the following) that have invaded the living body. One of them is called natural immune response system, another acquired immune response system. The former is an exclusion mechanism comprising phagocytosis by phagocytes (polymorphonuclear leukocytes, monocytes, macrophages, etc.), attack by natural killer (NK) cells, and non-specific recognition such as opsonization of antigen by complements. The latter, acquired immune response system, is an exclusion mechanism by lymphocytes (mainly, T cells and B cells) that acquired the specificity to the antigen (namely, activated lymphocytes). B cells that acquired antigen specificity attack the antigen existing outside of the cells through production of antibodies specific to the antigen. T cells that acquired antigen specificity (namely, activated T cells) are classified into helper T cells and cytotoxic T cells (cytotoxic lymphocyte, CTL). The helper T cells regulate a differentiation of B cells and a production of antibodies, and destroy the antigen cooperating with phagocytes. The latter, CTLs attack virus-infected cells and so on by themselves (Experimental Medicine: SUPPLEMENT, xe2x80x9cBio Science Term Library, Immunityxe2x80x9d, Yodosha, pp.14-17 (1995)).
This acquisition of antigen specificity by T cells (namely, activation of T cells) is initiated through recognition by T cells the antigen presented by antigen-presenting cells (APC) such as macrophage, B cells, or dendritic cells. Antigen-presenting cells process the antigens so incorporated and present these processed antigens through binding them to major histocompatibility complex (MHC). T cells receive primary signal for activation of the cells (or acquisition of specificity) by recognizing the processed antigens presented by antigen-presenting cells through a complex between T cell receptor (TcR) and CD3 antigen existing on the surface of the cell membrane (TcR/CD3 complex).
However, the TcR/CD3 complex-mediated primary signal alone cannot activate T cells sufficiently and leads to unresponsiveness or clonal anergy, so that the cells can not react with any stimulation received thereafter. The autocrine of interleukin 2 (IL-2) is necessary for T cells to be activated, to be differentiated into antigen specific T cell clones, and to be proliferated. In clonal anergy, T cells are inactivated due to no production of IL-2 and such and no cell division. Namely, the activation of T cells accompanied by production of cytokines such as IL-2 requires the secondary signal following the first signal through TcR/CD3 complex. This secondary signal is called costimulatory signal.
T cells receive this secondary signal and transmit it into the cells by interacting (cell adhesion) with molecules other than MHC on antigen-presenting cells through molecules other than TcR/CD3 complex on the T cell surface. This secondary signal avoids cell anergy (clonal anergy) and activates the cells.
Although some part of the mechanism of the secondary signal transmission between antigen-presenting cells and lymphocytes such as T cells have not yet been elucidated in detail, studies so far have revealed that an important factor for the secondary signal transmission is the interaction of CD28 (also named Tp44, T44, or 9.3 antigen), which is a cell surface molecule expressed mainly on T cells and thymus cells, with CD80 (also named B7-1, B7, BB1, or B7/BB1), which is a cell surface molecule expressed on antigen-presenting cells (macrophages, monocytes, dendritic cells, etc.) and with CD86 (also named B7-2 or B70), which is also a cell surface molecule on antigen-presenting cells (namely, cell adhesion through the binding between these molecules). Moreover, it has been experimentally elucidated that the interaction of Cytolytic T lymphocyte associated antigen 4 (CTLA-4), whose expression is thought to be enhanced depending on the secondary signal, with the CD80 (B7-1) and CD86 (B7-2) (namely. cell adhesion through the binding between these molecules) also plays an important role in the regulation of T cell activation by the secondary signal. In other words, the regulation of T cell activation by the transmission of the secondary signal involves at least the interaction between CD28 and CD80/CD86, the enhancement of CTLA-4 expression, which is thought to depend on the interaction, and the interaction between CTLA-4 and CD80/CD86.
CD28 is known to be a costimulator molecule transmitting the secondary signal (costimulatory signal) required for the activation of T cells and for the avoidance of anergy. The secondary signal transmitted by binding this molecule to costimulator molecules, CD80 (B7-1) and CD86 (B7-2), on antigen-presenting cells (cell adhesion through the binding between these molecules), stabilizes mRNA of Th1-type cytokines and consequently promotes production by T cells of a large amount of Th1-type cytokines such as IL-2, IFNxcex3, and TNFxcex1. The expression of CTLA-4 is induced by the primary signal transmitted through TcR/CD3, and the expression is also enhanced by the secondary signal transmitted by the binding between CD28 and CD80. It is being revealed that CTLA-4 receives these signals to work to inhibit T cell function, which is contrary to the activation of T cells by the secondary signal transmitted by CD28.
Human CD28 and CTLA-4 are type I glycoproteins whose molecular weights are 44 kD and 41 to 43 kD, respectively. Both have an immunoglobulin-like domain, belong to the immunoglobulin superfamily, and have both function as a cell adhesion molecule and function as a signal transmission molecule.
Human CD28 forms a homodimer with a disulfide bond while CTLA-4 exists as a monomer. Both CD28 and CTLA-4 genes are located at xe2x80x9c2q33xe2x80x9d on human chromosome and xe2x80x9c1Cxe2x80x9d on mouse chromosome, and are composed of four (4) exons. Human CD28 and CTLA-4 are composed of 220 and 223 amino acids, respectively, including the leader sequences, and amino acid homology between them is 20 to 30%.
The ligands for CD28 and CTLA-4 are CD80 (B7-1) and CD86 (B7-2) in human and mice. CTLA-4 has about 20 times as high affinity to both ligands as CD28. It has been elucidated that the amino acid sequence structures xe2x80x9cMYPPPY (Met-Tyr-Pro-Pro-Pro-Tyr; SEQ ID NO:41)xe2x80x9d conserved through animal species is important for the binding of CD28 and CTLA-4 to CD80 (B7-1). It has also been reported that, when CD28 is stimulated, PI3 kinase (phosphoinositide 3 kinase, PI3K) associates with the phosphorylated tyrosine residue in a partial sequence xe2x80x9cYMNM (Tyr-Met-Asn-Met; SEQ ID NO:42)xe2x80x9d of CD28 and that CD28 plays an important role in intracellular signal transmission through this xe2x80x9cYxxMxe2x80x9d structure. Furthermore, it has been reported that CTLA-4 also has a sequence represented by xe2x80x9cYxxM,xe2x80x9d namely xe2x80x9cYVKM (Tyr-Val-Lys-Met; SEQ ID NO:43)xe2x80x9d in its cytoplasmic region and that, after being stimulated, SYP associates with this sequence.
CD28 is expressed specifically in thymocytes and peripheral blood T cells, and CTLA-4 is expressed specifically in activated T cells (Cell Engineering: SUPPLEMENT, xe2x80x9cHandbook of Adhesion Moleculexe2x80x9d, Shujunsha, pp.93-102 (1994); ibid. pp.120-136; Experimental Medicine: SUPPLEMENT, xe2x80x9cBIO SCIENCE Term Library, Immunityxe2x80x9d, Yodosha, pp.94-98 (1995); Experimental Medicine: SUPPLEMENT, xe2x80x9cBIO SCIENCE Term Library, Intracellular Signal Transductionxe2x80x9d, Yodosha, pp.58-59 (1997); Nihon Rinsho, Vol.55, No.6, pp.215-220 (1997)).
In the regulation of T cell function (the activation and the inhibition of function of T cells), the importance of interactions among multiple molecules such as costimulator molecules (CD28, CD80 (B7-1), CD86 (B7-2), etc.) and CTLA-4, which cooperates with them, has thus been recognized, and this has been drawn attention to the relationship between these molecules and diseases, and the treatment of diseases by regulating the function of these molecules.
As described above, although a living body activates its acquired immune response system against antigens that are foreign bodies to the living body (self), it also has immunological tolerance so as to show no immune response against its own component (autoantigen). If immunological tolerance breaks down by some reason, immune response to the autoantigen occurs, autoantigen-reactive T cells are induced by the same mechanism as mentioned above to fall into abnormal state of immunity, and various autoimmune diseases are caused.
In other words, since non-stimulated antigen presenting cells (APC) in normal tissues do not express costimulatory molecules when the immune system of a living body is normal, T cells are in the unresponsiveness state to maintain immunological tolerance even if autoantigen-reactive T cells, which reacts with autoantigen, exist. It has been suggested that in abnormal state of immunity, more autoantigen-reactive T cells are activated due to abnormal excess and continuous expression of costimulatory molecules to thereby cause autoimmune diseases.
From such viewpoints recently, many attempts to treat various autoimmune diseases by modulating the transmission of costimulatory signals, for example, the above-mentioned signal transmission between CD28/CTLA-4 and CD80/CD86, are proposed.
The results of such attempts have not yet clarified in detail the mechanism of the T cell activation by interaction between costimulatory molecules and the related molecules. Other unknown molecules may be involved in this mechanism.
Recently, there has been identified a novel co-stimulatory molecule like the above-described xe2x80x9cCD28xe2x80x9d and xe2x80x9cCTLA-4xe2x80x9d, which is thought to carry out the transduction of a second signal (co-stimulatory signal) essential for the activation of lymphocytes such as T cells, and functional regulation coupled with said signal of activated lymphocytes such as activated T cells. This molecule has been designated as AILIM (activation inducible lymphocyte immunomodulatory molecule) (in humans, mice and rats: Int. Immunol., Vol. 12, No. 1, p.51-55, 2000), also referred to as ICOS (inducible co-stimulator) (in humans: Nature, Vol. 397, No.6716, p.263-266, 1999)).
On the other hand, novel molecules celled B7h, B7RP-1, GL50 or LICOS which are ligands (AILIM ligands) interacting with this costimulatory transmission molecule AILIM (ICOS) have been identified very recently (Nature. Vol.402, No.6763, pp.827-832, 1999; Nature Medicine, Vol.5, No.12, pp.1365-1369, 1999; J. Immunology, Vol.164, pp.1653-1657, 2000; Curr. Biol., Vol.10, No.6, pp.333-336, 2000).
The identification of these two kinds of novel molecules, namely AILIM (ICOS) and B7RP-1 (B7h, GL50, LICOS), as the signal transduction pathway for the costimulatory signal essential for the above activation of lymphocytes such as T cells, and the control of the function of activated T cells, revealed that there is the novel third pathway by the interaction between AILIM (ICOS) and B7RP-1 (B7h, GL50, LICOS), besides the known first and second signal pathways which are already known transduction pathway between CD28 and CD80 (B7-1)/CD86 (B7-2), and that between CTLA4 and CD80 (B7-1)/CD 86 (B7-2).
Studies on the biological functions of these novel molecules, the function control of lymphocytes, such as T cells, through this third costimulatory signal transduction by the molecules, and the relationship between the novel signal transduction and diseases are in progress (J. Immunol., 166(1), pp.1, 2001; J. Immunol., 165(9), pp.5035, 2000; Biochem. Biophys. Res. Commun., 276(1), pp.335, 2000; Immunity, 13(1), pp.95, 2000; J. Exp. Med., 192(1), pp.53, 2000; Eur. J. Immunol., 30(4), pp.1040, 2000; WO 01/15732).
Specifically, an objective of the present invention is to reveal biological functions of the novel molecule AILIM, considered, like xe2x80x9cCD28xe2x80x9d and xe2x80x9cCTLA-4xe2x80x9d, as a molecule which transmits the secondary signal (costimulatory signal) essential for the activation of lymphocytes, such as T cells, and which controls the functions of activated lymphocytes, such as activated T cells, by working with the signal; to reveal relationships between the expression of AILIM and diseases; and to provide a method and a pharmaceutical which inhibit the development of the various diseases dependent on the expression pattern of AILIM or which treat the diseases by controlling the biological functions of the AILIM using the medical and pharmaceutical methods (for example, a drug such as a monoclonal antibody and a low molecular compound).
To achieve the above-described purposes, the present inventors have actively pursued studies on human antibodies (particularly human monoclonal antibodies) against mammalian AILIMs (particularly human AILIM), and as a result, by immunizing transgenic mice prepared using genetic recombination techniques so as to produce human antibodies with AILIM (specifically cell membrane fraction of cells expressing human AILIM), succeeded first in the world in preparing a variety of monoclonal antibodies which bind to human AILIM, particularly those which bind to human AILIM that regulate signal transduction mediated by human AILIM.
Since antibodies (particularly monoclonal antibodies) of this invention are derived from humans, they do not induce any severe immune rejection due to the immunogenicity against humans , HAMA (human anti-mouse antigenicity), in the host at all, which has been a big problem (side effect) in therapy using antibody pharmaceuticals comprising antibodies derived from non-human mammals such as mice, and thus dramatically enhancing the value of antibody as medicine.
Therefore, human antibodies (particularly human monoclonal antibodies) which bind to mammalian AILIMs (particularly human AILIM) of this invention and pharmaceutical compositions comprising said human antibodies (particularly human monoclonal antibodies) are useful as drugs to control, with no induction of immune rejection due to HAMA in the host, various physiological reactions related to the transduction of co-stimulatory signal to AILIM-expressing cells mediated by AILIM (for example, proliferation of AILIM-expressing cells, cytokine production by AILIM-expressing cells, immune cytolysis or apoptosis of AILIM-expressing cells, and activity to induce antibody-dependent cytotoxicity to AILIM-expressing cells, and so on), and/or are also useful as drugs to suppress and prevent development of symptoms and/or progress of various disorders related to the signal transduction mediated by said AILIM, and as medicine to treat or prevent said disorder.
Specifically, pharmaceutical compositions according to this invention are able to control (suppress or stimulate) proliferation of AILIM-expressing cells or production of cytokine (for example, interferon g or interleukin 4, etc.) by AILIM-expressing cells, thereby enabling suppression of various pathological conditions and treatment or prevention of various disorders caused by diverse physiological phenomena related to signal transduction mediated by AILIM.
Use of pharmaceutical compositions according to this invention enables suppression, prevention and/or treatment of, for example, various disorders (for example, rheumatoid arthritis, multiple sclerosis, autoimmune thyroiditis, allergic contact-type dermatitis, chronic inflammatory dermatosis such as lichen planus, systemic lupus erythematosus, insulin-dependent diabetes mellitus, psoriasis, etc.) classified into autoimmune or allergic disorders (particularly autoimmune disease and delayed allergy caused by cellular immunity); arthropathia (for example, rheumatoid arthritis (RA) and osteoarthritis (OA)), inflammation (e.g., hepatitis); graft versus host reaction (GVH reaction); graft versus host disease (GVHD); immune rejection accompanying transplantation (homoplasty or heteroplasty) of a tissue (tissues such as skin, cornea, bone, etc.) or organ (liver, heart, lung, kidney, pancreas, etc.); immune response triggered by a foreign antigen or autoantigen (for example, production of antibodies against said antigen, cell proliferation, production of cytokines); and disorders possibly caused by the abnormal intestinal immunity (specifically inflammatory intestinal disorders (particularly clone disease and ulcerative colitis) and alimentary allergy).
Furthermore, in the field of suppression/treatment of immune rejection accompanying transplantation of above-described tissues and organs, it is possible to augment the suppressive effect on transplant rejection of known immunosuppressant by using the pharmaceutical composition of this invention together with said drugs which have been utilized for suppression of immune rejection in such a transplantation treatment.
Moreover, the pharmaceutical composition of the present invention can be applied for treating or preventing, any inflammatory diseases to which various steroids are indicated as antiphlogistic.
The pharmaceutical composition of the present invention can be applied to inflammatory disease for example, inflammation accompanying various arthritis (for example, rheumatoid arthritis, osteoarthritis), pneumonia, hepatitis (including viral hepatitis), inflammation accompanying infectious diseases, inflammatory bowel diseases, intestinal enteritis, nephritis (inflammation accompanying glomerular nephritis, nephrofibrosis), gastritis, angiitis, pancreatitis, peritonitis, bronchitis, myocarditis, cerebritis, inflammation in postischemic reperfusion injury (myocardial ischemic reperfusion injury), inflammation attributed to immune rejection after transplantation of tissue and organ, burn, various skin inflammation (psoriasis, allergic contact-type dermatitis, lichen planus which is chronic inflammatory skin disease), inflammation in multiple organ failure, inflammation after operation of PTCA or PTCR, and inflammation accompanying arteriosclerosis, and autoimmune thyroiditis.
In addition, by using a method for identifying substances that bind to AILIM or AILIM ligand, which is one of the present inventions, it becomes possible to screen to select pharmaceuticals (chemical synthetic compounds or antibodies) with potential activity to treat various disorders by binding to AILIM or AILIM ligands to regulate signal transduction mediated by interaction of them.
Specifically, the present invention is the invention described from the following (1) to (108).
(1) A human antibody which binds to AILIM.
(2) The human antibody of (1), wherein said AILIM is derived from human.
(3) A human monoclonal antibody which binds to AILIM or a portion thereof.
(4) The human monoclonal antibody or a portion thereof of (3), wherein said AILIM is derived from human.
(5) The human monoclonal antibody or a portion thereof of (3) or (4), wherein said human monoclonal antibody has an activity to inhibit a signal transduction into a cell mediated by AILIM.
(6) The human monoclonal antibody or a portion thereof of (5), wherein said activity to inhibit a signal transduction is (a) or (b) of the followings:
(a) activity to inhibit proliferation of AILIM-expressing cells, or
(b) activity to inhibit cytokine production from AILIM-expressing cells.
(7) The human monoclonal antibody or a portion thereof of (6), wherein said cytokine is one of the cytokines produced by Th1-type or Th2-type T cell.
(8) The human monoclonal antibody or a portion thereof of (7), wherein said cytokine is interferon xcex3 or interleukin 4.
(9) The human monoclonal antibody or a portion thereof of (5), wherein said human monoclonal antibody has an activity to prevent mixed lymphocyte reaction.
(10) The human monoclonal antibody or a portion thereof of (3) or (4), wherein said human monoclonal antibody has an activity to induce signal transduction into a cell mediated by AILIM.
(11) The human monoclonal antibody or a portion thereof of (10), wherein said activity to induce signal transduction is (a) or (b) of the followings:
(a) activity to induce proliferation of AILIM-expressing cells, or
(b) activity to induce cytokine production from AILIM-expressing cells.
(12) The human monoclonal antibody or a portion thereof of (11), wherein said cytokine is one of the cytokines produced by Th1-type or Th2-type T cell.
(13) The human monoclonal antibody or a portion thereof of (12), wherein said cytokine is interferon xcex3 or interleukin 4.
(14) The human monoclonal antibody or a portion thereof of (3) or (4), wherein said human monoclonal antibody has an activity to induce antibody-dependent cytotoxicity to AILIM-expressing cells, and/or immune cytolysis or apoptosis of AILIM-expressing cells.
(15) The human monoclonal antibody or a portion thereof of (3) or (4), wherein the binding rate constant (ka) between said monoclonal antibody and AILIM is 1.0xc3x97103 (1/M.Sec) or more.
(16) The human monoclonal antibody or a portion thereof of (15), wherein said binding rate constant (ka) is 1.0xc3x97104 (1/M.Sec) or more.
(17) The human monoclonal antibody or a portion thereof of (16), wherein said binding rate constant (ka) is 1.0xc3x97105 (1/M.Sec) or more.
(18) The human monoclonal antibody or a portion thereof of (3) or (4), wherein the dissociation rate constant (kd) between said monoclonal antibody and AILIM is 1.0xc3x9710xe2x88x923 (1/Sec) or less.
(19) The human monoclonal antibody or a portion thereof of (18), wherein said dissociation rate constant (kd) is 1.0xc3x9710xe2x88x924 (1/Sec) or less.
(20) The human monoclonal antibody or a portion thereof of (19), wherein said dissociation rate constant (kd) is 1.0xc3x9710xe2x88x925 (1/Sec) or less.
(21) The human monoclonal antibody or a portion thereof of (3) or (4), wherein the dissociation constant (Kd) between said monoclonal antibody and AILIM is 1.0xc3x9710xe2x88x926 (M) or less.
(22) The human monoclonal antibody or a portion thereof of (21), wherein said dissociation constant (Kd) is 1.0xc3x9710xe2x88x927 (M) or less.
(23) The human monoclonal antibody or a portion thereof of (22), wherein said dissociation constant (Kd) is 1.0xc3x9710xe2x88x928 (M) or less.
(24) The human monoclonal antibody or a portion thereof of (23), wherein said dissociation constant (Kd) is 1.0xc3x9710xe2x88x929 (M) or less.
(25) The human monoclonal antibody or a portion thereof of (4), wherein a V region DNA encoding a heavy chain variable region of said human monoclonal antibody is derived from either the human immunoglobulin heavy chain V gene segment 1-02 or 3-13.
(26) The human monoclonal antibody or a portion thereof of (4), wherein a V region DNA encoding a light chain variable region of said human monoclonal antibody is derived from either the human immunoglobulin light chain V gene segment L5 or A27.
(27) The human monoclonal antibody or a portion thereof of (25) or (26), wherein a V region DNA encoding a heavy chain variable region of said human monoclonal antibody is derived from either the human immunoglobulin heavy chain V gene segment 1-02 or 3-13, and wherein a V region DNA encoding a light chain variable region of said human monoclonal antibody is derived from either the human immunoglobulin light chain V gene segment L5 or A27.
(28) The human monoclonal antibody or a portion thereof of (27), wherein the V region DNA encoding a heavy chain variable region of said human monoclonal antibody is derived from the human immunoglobulin heavy chain V gene segment 1-02, and the V region DNA encoding a light chain variable region of said human monoclonal antibody is derived from the human immunoglobulin light chain V gene segment L5.
(29) The human monoclonal antibody or a portion thereof of (27), wherein the V region DNA encoding a heavy chain variable region of said human monoclonal antibody is derived from the human immunoglobulin heavy chain V gene segment 3-13, and the V region DNA encoding a light chain variable region of said human monoclonal antibody is derived from the human immunoglobulin light chain V gene segment A27.
(30) The human monoclonal antibody or a portion thereof of (4), wherein a heavy chain variable region of said human monoclonal antibody has an amino acid sequence defined in any of the following (a) through (f):
(a) amino acid sequence comprising amino acids from position 20 through 117 of SEQ ID NO:28,
(b) amino acid sequence comprising amino acids from position 20 through 117 of SEQ ID NO:28 in which one or more amino acid residues are deleted or substituted, or to which one or more amino acid residues are inserted or added.
(c) amino acid sequence comprising amino acids from position 20 through 116 of SEQ ID NO:32,
(d) amino acid sequence comprising amino acids from position 20 through 116 of SEQ ID NO:32 in which one or more amino acid residues are deleted or substituted, or to which one or more amino acid residues are inserted or added.
(e) amino acid sequence comprising amino acids from position 20 through 116 of SEQ ID NO:36, or
(f) amino acid sequence comprising amino acids from position 20 through 116 of SEQ ID NO:36, in which one or more amino acid residues are deleted or substituted, or to which one or more amino acid residues are inserted or added.
(31) The human monoclonal antibody or a portion thereof of (4), wherein a heavy chain polypeptide of said human monoclonal antibody has an amino acid sequence defined in any of the following (a) through (f):
(a) amino acid sequence comprising amino acids from position 20 through 470 of SEQ ID NO:28,
(b) amino acid sequence comprising amino acids from position 20 through 470 of SEQ ID NO:28 in which one or more amino acid residues are deleted or substituted, or to which one or more amino acid residues are inserted or added.
(c) amino acid sequence comprising amino acids from position 20 through 470 of SEQ ID NO:32,
(d) amino acid sequence comprising amino acids from position 20 through 470 of SEQ ID NO:32 in which one or more amino acid residues are deleted or substituted, or to which one or more amino acid residues are inserted or added.
(e) amino acid sequence comprising amino acids from position 20 through 470 of SEQ ID NO:36, or
(f) amino acid sequence comprising amino acids from position 20 through 470 of SEQ ID NO:36 in which one or more amino acid residues are deleted or substituted, or to which one or more amino acid residues are inserted or added.
(32) The human monoclonal antibody or a portion thereof of (4), wherein a light chain variable region of said human monoclonal antibody has an amino acid sequence defined in any of the following (a) through (f):
(a) amino acid sequence comprising amino acids from position 23 through 116 of SEQ ID NO:30,
(b) amino acid sequence comprising amino acids from position 23 through 116 of SEQ ID NO:30 in which one or more amino acid residues are deleted or substituted, or to which one or more amino acid residues are inserted or added.
(c) amino acid sequence comprising amino acids from position 21 through 116 of SEQ ID NO:34,
(d) amino acid sequence comprising amino acids from position 21 through 116 of SEQ ID NO:34 in which one or more amino acid residues are deleted or substituted, or to which one or more amino acid residues are inserted or added.
(e) amino acid sequence comprising amino acids from position 21 through 116 of SEQ ID NO:38, or
(f) amino acid sequence comprising amino acids from position 21 through 116 of SEQ ID NO:38 in which one or more amino acid residues are deleted or substituted, or to which one or more amino acid residues are inserted or added.
(33) The human monoclonal antibody or a portion thereof of (4), wherein a light chain polypeptide of said human monoclonal antibody has an amino acid sequence defined in any of the following (a) through (f):
(a) amino acid sequence comprising amino acids from position 23 through 236 of SEQ ID NO:30,
(b) amino acid sequence comprising amino acids from position 23 through 236 of SEQ ID NO:30 in which one or more amino acid residues are deleted or substituted, or to which one or more amino acid residues are inserted or added.
(c) amino acid sequence comprising amino acids from position 21 through 236 of SEQ ID NO:34,
(d) amino acid sequence comprising amino acids from position 21 through 236 of SEQ ID NO:34 in which one or more amino acid residues are deleted or substituted, or to which one or more amino acid residues are inserted or added.
(e) amino acid sequence comprising amino acids from position 21 through 236 of SEQ ID NO:38, or
(f) amino acid sequence comprising amino acids from position 21 through 236 of SEQ ID NO:38 in which one or more amino acid residues are deleted or substituted, or to which one or more amino acid residues are inserted or added.
(34) The human monoclonal antibody or a portion thereof of (4), wherein said human monoclonal antibody has the following characteristics (a) and (b):
(a) a heavy chain variable region has an amino acid sequence comprising the amino acid sequence from amino acid 20 through 117 according to SEQ ID NO:28, and
(b) a light chain variable region has an amino acid sequence comprising the amino acid sequence from amino acid 23 through 116 according to SEQ ID NO:30.
(35) The human monoclonal antibody or a portion thereof of (4), wherein said human monoclonal antibody has the following characteristics (a) and (b):
(a) a heavy chain polypeptide has an amino acid sequence from amino acid 20 through 470 according to SEQ ID NO:28, and
(b) a light chain polypeptide has an amino acid sequence from amino acid 23 through 236 according to SEQ ID NO:30.
(36) The human monoclonal antibody or a portion thereof of (4), wherein said human monoclonal antibody has the following characteristics (a) and (b):
(a) a heavy chain variable region has an amino acid sequence comprising the amino acid sequence from amino acid 20 through 116 according to SEQ ID NO:32, and
(b) a light chain variable region has an amino acid sequence comprising the amino acid sequence from amino acid 21 through 116 according to SEQ ID NO:34.
(37) The human monoclonal antibody or a portion thereof of (4), wherein said human monoclonal antibody has the following characteristics (a) and (b):
(a) a heavy chain polypeptide has an amino acid sequence comprising the amino acid sequence from amino acid 20 through 470 according to SEQ ID NO:32, and
(b) a light chain polypeptide has an amino acid sequence comprising the amino acid sequence from amino acid 21 through 236 according to SEQ ID NO:34.
(38) The human monoclonal antibody or a portion thereof of (4), wherein said human monoclonal antibody has the following characteristics (a) and (b):
(a) a heavy chain variable region has an amino acid sequence comprising the amino acid sequence from amino acid 20 through 116 according to SEQ ID NO:36, and
(b) a light chain variable region has an amino acid sequence comprising the amino acid sequence from amino acid 21 through 116 according to SEQ ID NO:38.
(39) The human monoclonal antibody or a portion thereof of (4), wherein said human monoclonal antibody has the following characteristics (a) and (b):
(a) a heavy chain polypeptide has an amino acid sequence comprising the amino acid sequence from amino acid 20 through 470 according to SEQ ID NO:36, and
(b) a light chain polypeptide has an amino acid sequence comprising the amino acid sequence from amino acid 21 through 236 according to SEQ ID NO:38.
(40) The human monoclonal antibody or a portion thereof of any one of (3) through (29), wherein said human monoclonal antibody is a monoclonal antibody derived from a transgenic non-human mammal capable of producing human antibodies.
(41) The human monoclonal antibody or a portion thereof of (40), wherein said human monoclonal antibody is obtained by immunizing transgenic non-human mammal capable of producing human antibody with AILIM-expressing cells, membrane fractions derived from said cells, whole molecules constituting AILIM or a portion thereof, or genes encoding AILIM or a portion thereof.
(42) The human monoclonal antibody or a portion thereof of (40) or (41), wherein said transgenic non-human mammal is a transgenic mouse.
(43) A DNA or a portion thereof encoding a polypeptide selected from the group consisting of (a) through (f) below:
(a) a polypeptide comprising the amino acid sequence from amino acid 20 through 117 according to SEQ ID NO:28,
(b) a polypeptide comprising the amino acid sequence from amino acid 23 through 116 according to SEQ ID NO:30,
(c) a polypeptide comprising the amino acid sequence from amino acid 20 through 116 according to SEQ ID NO:32,
(d) a polypeptide comprising the amino acid sequence from amino acid 21 through 116 according to SEQ ID NO:34,
(e) a polypeptide comprising the amino acid sequence from amino acid 20 through 116 according to SEQ ID NO:36, and
(f) a polypeptide comprising the amino acid sequence from amino acid 21 through 116 according to SEQ ID NO:38.
(44) A DNA or a portion thereof encoding a polypeptide selected from the group consisting of (a) through (f) below:
(a) a polypeptide comprising the amino acid sequence from amino acids 20 through 470 according to SEQ ID NO:28,
(b) a polypeptide comprising the amino acid sequence from amino acids 23 through 236 according to SEQ ID NO:30,
(c) a polypeptide comprising the amino acid sequence from amino acids 20 through 470 according to SEQ ID NO:32,
(d) a polypeptide comprising the amino acid sequence from amino acids 21 through 236 according to SEQ ID NO:34,
(e) a polypeptide comprising the amino acid sequence from amino acids 20 through 470 according to SEQ ID NO:36, and
(f) a polypeptide comprising the amino acid sequence from amino acids 21 through 236 according to SEQ ID NO:38.
(45) A DNA or a portion thereof selected from the group consisting of (a) through (f) below:
(a) a DNA comprising the nucleotide sequence from nucleotides 126 through 419 according to SEQ ID NO:27,
(b) a DNA comprising the nucleotide sequence from nucleotides 105 through 386 according to SEQ ID NO:29,
(c) a DNA comprising the nucleotide sequence from nucleotides 151 through 441 according to SEQ ID NO:31,
(d) a DNA comprising the nucleotide sequence from nucleotides 88 through 375 according to SEQ ID NO:33,
(e) a DNA comprising the nucleotide sequence from nucleotides 153 through 443 according to SEQ ID NO:35, and
(f) a DNA comprising the nucleotide sequence from nucleotides 93 through 380 according to SEQ ID NO:37.
(46) A DNA or a portion thereof selected from a group consisting of (a) through (f) below:
(a) a DNA comprising the nucleotide sequence from nucleotides 69 through 1481 according to SEQ ID NO:27,
(b) a DNA comprising the nucleotide sequence from nucleotides 39 through 749 according to SEQ ID NO:29,
(c) a DNA comprising the nucleotide sequence from nucleotides 94 through 1506 defined in SEQ ID NO:31,
(d) a DNA comprising the nucleotide sequence from nucleotides 28 through 738 according to SEQ ID NO:33,
(e) a DNA comprising the nucleotide sequence from nucleotides 96 through 1508 according to SEQ ID NO:35, and
(f) a DNA comprising the nucleotide sequence from nucleotides 33 through 743 according to SEQ ID NO:37.
(47) A vector comprising the DNA of any one of (43) through (46).
(48) The vector of (47) comprising a DNA according to any of the following (a) through (c):
(a) a DNA comprising the nucleotide sequence from nucleotides 126 through 419 according to SEQ ID NO:27,
(b) a DNA comprising the nucleotide sequence from nucleotides 151 through 441 according to SEQ ID NO:31, or
(c) a DNA comprising the nucleotide sequence from nucleotides 153 through 443 according to SEQ ID NO:35.
(49) The vector of (47) comprising a DNA according to any of the following (a) through (c):
(a) a DNA comprising the nucleotide sequence from nucleotides 69 through 1481 according to SEQ ID NO:27,
(b) a DNA comprising the nucleotide sequence from nucleotides 94 through 1506 according to SEQ ID NO:31, or
(c) a DNA comprising the nucleotide sequence from nucleotides 96 through 1508 according to SEQ ID NO:35.
(50) The vector of (47) comprising a DNA according to any of the following (a) through (c):
(a) a DNA comprising the nucleotide sequence from nucleotides 105 through 386 according to SEQ ID NO:29,
(b) a DNA comprising the nucleotide sequence from nucleotides 88 through 375 according to SEQ ID NO:33, or
(c) a DNA comprising the nucleotide sequence from nucleotides 93 through 380 according to SEQ ID NO:37.
(51) The vector of (47) comprising a DNA according to any of the following (a) through (c):
(a) a DNA comprising the nucleotide sequence from nucleotides 39 through 749 according to SEQ ID NO:29,
(b) a DNA comprising the nucleotide sequence from nucleotides 28 through 738 according to SEQ ID NO:33, or
(c) a DNA comprising the nucleotide sequence from nucleotides 33 through 743 according to SEQ ID NO:37.
(52) The vector of (47) comprising a DNA according to the following (a) and (b):
(a) a DNA comprising the nucleotide sequence from nucleotides 126 through 419 according to SEQ ID NO:27, and
(b) a DNA comprising the nucleotide sequence from nucleotides 105 through 386 according to SEQ ID NO:29.
(53) The vector of (47) comprising a DNA according to the following (a) and (b):
(a) a DNA comprising the nucleotide sequence from nucleotides 69 through 1481 according to SEQ ID NO:27, and
(b) a DNA comprising the nucleotide sequence from nucleotides 39 through 749 according to SEQ ID NO:29.
(54) The vector of (47) comprising a DNA according to the following (a) and (b):
(a) a DNA comprising the nucleotide sequence from nucleotides 151 through 441 according to SEQ ID NO:31, and
(b) a DNA comprising the nucleotide sequence from nucleotides 88 through 375 according to SEQ ID NO:33.
(55) The vector of (47) comprising a DNA according to the following (a) and (b):
(a) a DNA comprising the nucleotide sequence from nucleotides 94 through 1506 according to SEQ ID NO:31, and
(b) a DNA comprising the nucleotide sequence from nucleotides 28 through 738 according to SEQ ID NO:33.
(56) The vector of (47) comprising a DNA according to the following (a) and (b):
(a) a DNA comprising the nucleotide sequence from nucleotides 153 through 443 according to SEQ ID NO:35, and
(b) a DNA comprising the nucleotide sequence from nucleotides 93 through 380 according to SEQ ID NO:37.
(57) The vector of (47) comprising a DNA according to the following (a) and (b):
(a) a DNA comprising the nucleotide sequence from nucleotides 96 through 1508 according to SEQ ID NO:35, and
(b) a DNA comprising the nucleotide sequence from nucleotides 33 through 743 according to SEQ ID NO:37.
(58) A cell producing a human monoclonal antibody of any one of (3) through (42).
(59) The cell of (58), wherein said cell is a fused cell obtained by fusing B cell, derived from a mammal capable of producing said human monoclonal antibody, and myeloma cell derived from a mammal.
(60) A genetic recombinant host transformed by transferring a DNA described below in (a) or a vector comprising said DNA, a DNA described below in (b) or a vector comprising said DNA, or both DNAs described below in (a) and (b) or a vector comprising both of said DNAs:
(a) a DNA encoding a heavy chain polypeptide or a portion thereof of a monoclonal antibody which binds to human AILIM; or
(b) a DNA encoding a light chain polypeptide or a portion thereof of a monoclonal antibody which binds to human AILIM.
(61) The genetic recombinant host of (60), wherein said monoclonal antibody is a human monoclonal antibody.
(62) The genetic recombinant host of (60) or (61), wherein said host is a mammalian cell.
(63) The genetic recombinant host of (60) or (61), wherein said host is a mammalian fertilized egg.
(64) The genetic recombinant host of any one of (60) through (63), wherein said heavy chain polypeptide is one of the heavy chain polypeptides selected from the group consisting of the following (a) through (c):
(a) a heavy chain polypeptide comprising the amino acid sequence from amino acids 20 through 117 according to SEQ ID NO:28,
(b) a heavy chain polypeptide comprising the amino acid sequence from amino acids 20 through 116 according to SEQ ID NO:32, and
(c) a heavy chain polypeptide comprising the amino acid sequence from amino acids 20 through 116 according to SEQ ID NO:36.
(65) The genetic recombinant host of any one of (60) through (63), wherein said heavy chain polypeptide is one of the heavy chain polypeptide selected from the group consisting of the following (a) through (c):
(a) a heavy chain polypeptide comprising the amino acid sequence from amino acids 20 through 470 according to SEQ ID NO:28,
(b) a heavy chain polypeptide comprising the amino acid sequence from amino acids 20 through 470 according to SEQ ID NO:32, and
(c) a heavy chain polypeptide comprising the amino acid sequence from amino acids 20 through 470 according to SEQ ID NO:36.
(66) The genetic recombinant host of any one of (60) through (63), wherein said light chain polypeptide is one of the light chain polypeptide selected from the group consisting of the following (a) through (c):
(a) a heavy chain polypeptide comprising the amino acid sequence from amino acids 23 through 116 according to SEQ ID NO:30,
(b) a heavy chain polypeptide comprising the amino acid sequence from amino acids 21 through 116 according to SEQ ID NO:34, and
(c) a heavy chain polypeptide comprising the amino acid sequence from amino acids 21 through 116 according to SEQ ID NO:38.
(67) The genetic recombinant host of any one of (60) through (63), wherein said light chain polypeptide is one of the light chain polypeptide selected from the group consisting of the following (a) through (c):
(a) a light chain polypeptide comprising the amino acid sequence from amino acids 23 through 236 according to SEQ ID NO:30,
(b) a light chain polypeptide comprising the amino acid sequence from amino acids 21 through 236 according to SEQ ID NO:34, and
(c) a light chain polypeptide comprising the amino acid sequence from amino acids 21 through 236 according to SEQ ID NO:38.
(68) The genetic recombinant host of any one of (60) through (63), wherein said heavy chain and light chain polypeptides are those defined below in (a) and (b), respectively:
(a) a heavy chain polypeptide comprising the amino acid sequence from amino acids 20 through 117 according to SEQ ID NO:28, and
(b) a light chain polypeptide comprising the amino acid sequence from amino acids 23 through 116 according to SEQ ID NO:30.
(69) The genetic recombinant host of any one of (60) through (63), wherein said heavy chain and light chain polypeptides are those defined below in (a) and (b), respectively:
(a) a heavy chain polypeptide comprising the amino acid sequence from amino acids 20 through 470 according to SEQ ID NO:28, and
(b) a light chain polypeptide comprising the amino acid sequence from amino acids 23 through 236 according to SEQ ID NO:30.
(70) The genetic recombinant host of any one of (60) through (63), wherein said heavy chain and light chain polypeptides are those defined below in (a) and (b), respectively:
(a) a heavy chain polypeptide comprising the amino acid sequence from amino acids 20 through 116 according to SEQ ID NO: 32, and
(b) a light chain polypeptide comprising the amino acid sequence from amino acids 21 through 116 according to SEQ ID NO:34.
(71) The genetic recombinant host of any one of (60) through (63), wherein said heavy chain and light chain polypeptides are those defined below in (a) and (b), respectively:
(a) a heavy chain polypeptide comprising the amino acid sequence from amino acids 20 through 470 according to SEQ ID NO:32, and
(b) a light chain polypeptide comprising the amino acid sequence from amino acids 21 through 236 according to SEQ ID NO:34.
(72) The genetic recombinant host of any one of (60) through (63), wherein said heavy chain and light chain polypeptides are those defined below in (a) and (b), respectively:
(a) a heavy chain polypeptide comprising the amino acid sequence from amino acids 20 through 116 according to SEQ ID NO:36, and
(b) a light chain polypeptide comprising the amino acid sequence from amino acids 21 through 116 according to SEQ ID NO:38.
(73) The genetic recombinant host of any one of (60) through (63), wherein said heavy chain and light chain polypeptides are those defined below in (a) and (b), respectively:
(a) a heavy chain polypeptide comprising the amino acid sequence from amino acids 20 through 470 according to SEQ ID NO:36, and
(b) a light chain polypeptide comprising the amino acid sequence from amino acids 21 through 236 according to SEQ ID NO:38.
(74) The genetic recombinant host of any one of (60) through (63), wherein the DNA encoding said heavy chain polypeptide is a DNA defined in any of following (a) through (c):
(a) a DNA comprising the nucleotide sequence from nucleotides 126 through 419 according to SEQ ID NO:27,
(b) a DNA comprising the nucleotide sequence from nucleotides 151 through 441 according to SEQ ID NO:31, and
(c) a DNA comprising the nucleotide sequence from nucleotides 153 through 443 according to SEQ ID NO:35.
(75) The genetic recombinant host of any one of (60) through (63), wherein the DNA encoding said heavy chain polypeptide is a DNA defined in any of following (a) through (c):
(a) a DNA comprising the nucleotide sequence from nucleotides 69 through 1481 according to SEQ ID NO:27,
(b) a DNA comprising the nucleotide sequence from nucleotides 94 through 1506 according to SEQ ID NO:31, and
(c) a DNA comprising the nucleotide sequence from nucleotides 96 through 1508 according to SEQ ID NO:35.
(76) The genetic recombinant host of any one of (60) through (63), wherein the DNA encoding said light chain polypeptide is a DNA defined in any of following (a) through (c):
(a) a DNA comprising the nucleotide sequence from nucleotides 105 through 386 according to SEQ ID NO:29,
(b) a DNA comprising the nucleotide sequence from nucleotides 88 through 375 according to SEQ ID NO:33, and
(c) a DNA comprising the nucleotide sequence from nucleotides 93 through 380 according to SEQ ID NO:37.
(77) The genetic recombinant host of any one of (60) through (63), wherein the DNA encoding said light chain polypeptide is a DNA as defined in any of following (a) through (c):
(a) a DNA comprising the nucleotide sequence from nucleotides 39 through 749 according to SEQ ID NO:29,
(b) a DNA comprising the nucleotide sequence from nucleotides 28 through 738 according to SEQ ID NO:33, and
(c) a DNA comprising the nucleotide sequence from nucleotides 33 through 743 according to SEQ ID NO:37.
(78) The genetic recombinant host of any one of (60) through (63), wherein the DNA encoding said heavy chain polypeptide is a DNA described below in (a), and the DNA encoding said light chain polypeptide is a DNA as described below in (b):
(a) a DNA comprising the nucleotide sequence from nucleotides 126 through 419 according to SEQ ID NO:27, and
(b) a DNA comprising the nucleotide sequence from nucleotides 105 through 386 according to SEQ ID NO:29.
(79) The genetic recombinant host of any one of (60) through (63), wherein the DNA encoding said heavy chain polypeptide is the DNA described below in (a), and the DNA encoding said light chain polypeptide is the DNA described below in (b):
(a) a DNA comprising the nucleotide sequence from nucleotides 69 through 1481 according to SEQ ID NO:27, and
(b) a DNA comprising the nucleotide sequence from nucleotides 39 through 749 according to SEQ ID NO:29.
(80) The genetic recombinant host of any one of (60) through (63), wherein the DNA encoding said heavy chain polypeptide is the DNA described below in (a), and the DNA encoding said light chain polypeptide is the DNA described below in (b):
(a) a DNA comprising the nucleotide sequence from nucleotides 151 through 441 according to SEQ ID NO:31, and
(b) a DNA comprising the nucleotide sequence from nucleotides 88 through 375 V SEQ ID NO:33.
(81) The genetic recombinant host of any one of (60) through (63), wherein the DNA encoding said heavy chain polypeptide is the DNA described below in (a), and the DNA encoding said light chain polypeptide is the DNA described below in (b):
(a) a DNA comprising the nucleotide sequence from nucleotides 94 through 1506 according to SEQ ID NO:31, and
(b) a DNA comprising the nucleotide sequence from nucleotides 28 through 738 according to SEQ ID NO:33.
(82) The genetic recombinant host of any one of (60) through (63), wherein the DNA encoding said heavy chain polypeptide is the DNA described below in (a), and the DNA encoding said light chain polypeptide is the DNA described below in (b):
(a) a DNA comprising the nucleotide sequence from nucleotides 153 through 443 according to SEQ ID NO:35, and
(b) a DNA comprising the nucleotide sequence from nucleotides 93 through 380 according to SEQ ID NO:37.
(83) The genetic recombinant host of any one of (60) through (63), wherein the DNA encoding said heavy chain polypeptide is the DNA described below in (a), and the DNA encoding said light chain polypeptide is the DNA described below in (b):
(a) a DNA comprising the nucleotide sequence from nucleotides 96 through 1508 according to SEQ ID NO:35, and
(b) a DNA comprising the nucleotide sequence from nucleotides 33 through 743 according to SEQ ID NO:37.
(84) A human monoclonal antibody or a portion thereof produced by a genetic recombinant host (provided excluding the case where said host is a fertilized egg) of any one of (60) through (62), or of any one of (64) through (83).
(85) A pharmaceutical composition comprising the human antibody of (1) or (2), and a pharmaceutically acceptable carrier.
(86) A pharmaceutical composition comprising the human monoclonal antibody or a portion thereof of any one of (3) to (42), and a pharmaceutically acceptable carrier.
(87) A pharmaceutical composition comprising a human monoclonal antibody or a portion thereof of (84), and a pharmaceutically acceptable carrier.
(88) The pharmaceutical composition of any one of (85) through (87), wherein said pharmaceutical composition is used to inhibit signal transduction into the cell mediated by AILIM.
(89) The pharmaceutical composition of any one of (85) through (87), wherein said pharmaceutical composition is used to prevent proliferation of AILIM-expressing cells.
(90) The pharmaceutical composition of any one of (85) through (87), wherein said pharmaceutical composition is used to prevent production of a cytokine from AILIM-expressing cells.
(91) The pharmaceutical composition of any one of (85) through (87), wherein said pharmaceutical composition is used to induce signal transduction into a cell mediated by AILIM.
(92) The pharmaceutical composition of any one of (85) through (87), wherein said pharmaceutical composition is used to induce proliferation of AILIM-expressing cells.
(93) The pharmaceutical composition of any one of (85) through (87), wherein said pharmaceutical composition is used to induce production of a cytokine from AILIM-expressing cells.
(94) The pharmaceutical composition of any one of (85) through (87), wherein said pharmaceutical composition is used to induce antibody-dependent cytotoxicity against AILIM-expressing cells, and/or immune cytolysis or apoptosis of AILIM-expressing cells.
(95) A pharmaceutical composition for preventing, treating, or prophylaxis of delayed type allergy, comprising a substance having an activity in modulating signal transduction mediated by AILIM, and a pharmaceutically acceptable carrier.
(96) The pharmaceutical composition of (95), wherein the substance is a protein substance.
(97) The pharmaceutical composition of (96), wherein the protein substance is selected from the group consisting of:
a) an antibody which binds to AILIM or a portion thereof;
b) a polypeptide comprising the whole or a portion of an extracellular region of AILIM;
c) a fusion polypeptide comprising the whole or a portion of an extracellular region of AILIM, and the whole or a portion of a constant region of immunoglobulin heavy chain; and
d) a polypeptide which binds to AILIM.
(98) The pharmaceutical composition of (97), wherein said antibody that binds to AILIM is the human antibody of (1) or (2).
(99) The pharmaceutical composition of (97), wherein said antibody that binds to AILIM is the human monoclonal antibody of any one of (3) through (42).
(100) The pharmaceutical composition of (97), wherein said antibody against AILIM is the human monoclonal antibody of (84).
(101) The pharmaceutical composition of (95), wherein the substance is a non-protein substance.
(102) The pharmaceutical composition of (101), wherein the non-protein substance is DNA, RNA, or a chemically synthesized compound.
(103) A method for identifying substances that bind to AILIM or AILIM ligand comprising the following processes:
(a) preparing an insoluble carrier on which the entire extracellular region of AILIM or a portion thereof is immobilized;
(b) preparing a polypeptide comprising the whole extracellular region of AILIM ligand or a portion thereof labeled with a labeling material that emit a detectable signal;
(c) reacting the insoluble carrier in process(a) with the polypeptide in process (b);
(d) reacting the insoluble carrier of process (a), the polypeptide of process (b) and said substance to each other in any arbitrary orders;
(e) detecting the signal emitted from said labeling material contained in the complex produced in process (c), and the signal emitted from said labeling material contained in the complex produced in process (d), respectively; and
(f) comparing the magnitude of each of signals detected in process (e).
(104) A method for identifying substances that bind to AILIM or AILIM ligand comprising the following processes:
(a) preparing an insoluble carrier on which the entire extracellular region of AILIM ligand or a portion thereof is immobilized;
(b) preparing a polypeptide comprising the whole extracellular region of AILIM or a portion thereof labeled with a labeling material that emit a detectable signal;
(c) reacting the insoluble carrier in process (a) with the polypeptide in process (b);
(d) reacting the insoluble carrier of process (a), the polypeptide of process (b) and said substance to each other in any arbitrary orders;
(e) detecting the signal emitted from said labeling material contained in the complex produced in process (c), and the signal emitted from said labeling material contained in the complex produced in process (d), respectively; and
(f) comparing the magnitude of each of signals detected in process (e).
(105) The method of (103) or (104), wherein said polypeptide comprising the whole extracellular region of AILIM or a portion thereof is a fusion polypeptide comprising a polypeptide, comprising the whole extracellular region of AILIM or a portion thereof, and the whole constant region of immunoglobulin heavy chain or a portion thereof.
(106) The method of (103) or (104), wherein said polypeptide comprising the whole extracellular region of AILIM ligand or a portion thereof is a fusion polypeptide comprising a polypeptide, comprising the whole extracellular region of AILIM ligand or a portion thereof, and the whole constant region of immunoglobulin heavy chain or a portion thereof.
(107) The method of any one of (103) through (106), wherein said AILIM is a human AILIM.
(108) The method of any one of (103) through (107), wherein said AILIM ligand is a human AILIM ligand.